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Objective: To evaluate the clinical effects of low- and intermediate-dose factor Ⅷ (F Ⅷ) prophylaxis in Chinese adult patients with severe hemophilia A. Methods: Thirty adult patients with severe hemophilia A who received low- (n=20) /intermediate-dose (n=10) F Ⅷ prophylaxis at Nanjing Drum Tower Hospital affiliated with Nanjing University Medical College were included in the study. The annual bleeding rate (ABR), annual joint bleeding rate (AJBR), number of target joints, functional independence score of hemophilia (FISH), quality of life score, and health status score (SF-36) before and after preventive treatment were retrospectively analyzed and compared. Results: The median follow-up was 48 months. Compared with on-demand treatment, low- and intermediate-dose prophylaxis significantly reduced ABR, AJBR, and the number of target joints (P<0.05) ; the improvement in the intermediate-dose prophylaxis group was better than that in the low-dose prophylaxis group (P<0.05). Compared with on-demand treatment, the FISH score, quality of life score, and SF-36 score significantly improved in both groups (P<0.05), but there was no significant difference between the two groups (P>0.05) . Conclusion: In Chinese adults with severe hemophilia A, low- and intermediate-dose prophylaxis can significantly reduce bleeding frequency, delay the progression of joint lesions, and improve the quality of life of patients as compared with on-demand treatment. The improvement in clinical bleeding was better with intermediate-dose prophylaxis than low-dose prophylaxis.
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Humanos , Hemofilia A/tratamento farmacológico , Fator VIII/uso terapêutico , Qualidade de Vida , Estudos Retrospectivos , Hemartrose/prevenção & controle , Hemorragia/tratamento farmacológicoRESUMO
OBJECTIVE@#To analyze the clinical characteristics and long-term prognosis of patients with primary bone lymphoma (PBL).@*METHODS@#The clinical data of 21 patients with PBL treated in our center from 2005 to 2018 were analyzed retrospectively, the clinical characteristics and the factors affecting prognosis of the patients were analyzed.@*RESULTS@#The median age of all the 21 newly diagnosed PBL patients was 40(12-71) years old. Ostealgia was the initial symptom in most of the patients (19/21,90.5%). 42.9%(9/21) of the patients showed single bone lesion only. 571% (12/21) of the patients showed diffuse large B cell lymphoma. 28.6% (6/21) of the patients showed anaplastic large cell lymphoma and 9.5% (2/21) of the patients showed T cell lymphoblastic lymphoma. All the patients received chemotherapy (CHOP or CHOP like regimen, 33.3% plus rituximab) with or without radiotherapy and/or autologous hematopoietic stem cell transplantation (ASCT). 18 patients achieved clinical remission (including 15 for CR and 3 for PR). The median follow-up time was 48 months. The 5-year overall survival rate and progression-free survival rate of the patients were was 67.5% and 63.7%, respectively. The single factors analysis showed that ASCT was the important prognostic factor of PFS, while the single or multiple bone lesion was the factors affecting OS of the patients. There were no statistical differences with the effects of age, sex, stage, ECOG score, LDH level, B symptoms and radiotherapy for the prognosis of patients.@*CONCLUSION@#Diffuse large B cell lymphoma is the most common pathological type of PBL. Chemotherapy is the main treatment, which can be combined with radiotherapy and/or ASCT. The ASCT and the number of bone lesion are the factors for long time survival of the patients.
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Adulto , Idoso , Humanos , Pessoa de Meia-Idade , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Ciclofosfamida , Intervalo Livre de Doença , Doxorrubicina , Linfoma Difuso de Grandes Células B/tratamento farmacológico , Prednisona , Prognóstico , Estudos Retrospectivos , Rituximab/uso terapêutico , Transplante Autólogo , VincristinaRESUMO
OBJECTIVE@#To study the regulatory effect of deubiquitinase MYSM1 on differentiation of B cells to plasma cells.@*METHODS@#The interfering and overexpression plasmids of MYSM1 were constructed and then the corresponding lentiviruses were packaged. Human CD19 B cells were isolated from human peripheral blood with Miltenyi B cell isolation kit. Purified CD19 B cells were transduced with lentiviruses and then treated with LPS, the CD138 expression was detected by flow cytometry. The expression of transcription factor was determined by quantitative PCR.@*RESULTS@#The differentiation of B cells to plasma cells was enhanced after interfering in MYSM1 expression. Quantitative PCR showed that mRNA levels of Pax5 and Bach2 in cells with interfering in MYSM1 were much lower than their counterpart (P<0.01), and mRNA levels of Prdm1 and Xbp1 in cells with interfering in MYSM1 were much higher than their counterpart (P<0.01). On the contrary, the differentiation of B cells to plasma cells was inhibited after the overexpression of MYSM1. Quantitative PCR showed that mRNA levels of Pax5 and Bach2 in cells with MYSM1 overexpression were higher than those in control cells (P<0.01), and mRNA levels of Prdm1 and Xbp1 in cells with MYSM1 overexpression were much lower than those in their counterpart (P<0.01).@*CONCLUSION@#MYSM1 negatively regulates differentiation of human B cells to plasma cells.
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Humanos , Linfócitos B , Diferenciação Celular , Proteínas de Ligação a DNA , Genética , Enzimas Desubiquitinantes , Plasmócitos , Fatores de Transcrição , GenéticaRESUMO
Background@#Prospective real-life data on the safety and effectiveness of rituximab in Chinese patients with diffuse large B-cell lymphoma (DLBCL) or follicular lymphoma (FL) are limited. This real-world study aimed to evaluate long-term safety and effectiveness outcomes of rituximab plus chemotherapy (R-chemo) as first-line treatment in Chinese patients with DLBCL or FL. Hepatitis B virus (HBV) reactivation management was also investigated.@*Methods@#A prospective, multicenter, single-arm, noninterventional study of previously untreated CD20-positive DLBCL or FL patients receiving first-line R-chemo treatment at 24 centers in China was conducted between January 17, 2011 and October 31, 2016. Enrolled patients underwent safety and effectiveness assessments after the last rituximab dose and were followed up for 3 years. Effectiveness endpoints included progression-free survival (PFS) and overall survival (OS). Safety endpoints were adverse events (AEs), serious AEs, drug-related AEs, and AEs of special interest. We also reported data on the incidence of HBV reactivation.@*Results@#In total, 283 previously untreated CD20-positive DLBCL and 31 FL patients from 24 centers were enrolled. Three-year PFS was 59% (95% confidence interval [CI]: 50-67%) for DLBCL patients and 46% (95% CI: 20-69%) for FL patients. For DLBCL patients, multivariate analyses showed that PFS was not associated with international prognostic index, tumor maximum diameter, HBV infection status, or number of rituximab treatment cycles, and OS was only associated with age >60 years (P < 0.05). R-chemo was well tolerated. The incidence of HBV reactivation in hepatitis B surface antigen (HBsAg)-positive and HBsAg-negative/hepatitis B core antibody-positive patients was 13% (3/24) and 4% (3/69), respectively.@*Conclusions@#R-chemo is effective and safe in real-world clinical practice as first-line treatment for DLBCL and FL in China, and that HBV reactivation during R-chemo is manageable with preventive measures and treatment.@*Trial Registration@#ClinicalTrials.gov, NCT01340443; https://clinicaltrials.gov/ct2/show/NCT01340443.
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Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Protocolos de Quimioterapia Combinada Antineoplásica , Usos Terapêuticos , China , Ciclofosfamida , Doxorrubicina , Seguimentos , Linfoma Folicular , Tratamento Farmacológico , Linfoma Difuso de Grandes Células B , Tratamento Farmacológico , Estudos Prospectivos , Rituximab , Usos Terapêuticos , VincristinaRESUMO
BACKGROUND: miR-15b plays an important role in the initiation and development of tumors, based on which, we speculate that miR-15b may be involved in the migration and invasion of glioma stem cells (GSCs). However, there is no relevant report and the mechanism of action is also unclear. OBJECTIVE: To identify the effects of miR-15b on the migration and invasion of GSCs and the mechanisms involved in this process. METHODS: Quantitative PCR was performed to evaluate the expression of miR-15b in the gliomas tissues, normal brain tissues, GSCs and non-GSCs. After knockdown of ATP-binding cassette superfamily G member 2 (ABCG2) by ABCG2 specific siRNA, Transwell assay was performed to determine the effect of ABCG2 on GSCs migration and invasion. Additionally, the GSCs were transfected with miR-15b mimics or inhibitor to up-regulate or down-regulate the expression of miR-15b. At 48 hours after transfection, Transwell assay was used to detect the effect of miR-15b on GSCs migration and invasion; ELISA and gelatin zymography assays were performed to determine the matrix metalloproteinase-2/-9 (MMP-2/-9) expression and activity after treatment with miR-15b. CD133-positive or non-CD133-positive cells were directly injected subcutaneously into nude mice. Tumor formation was observed within 30 days after injection. RESULTS AND CONCLUSION: miR-15b was significantly down-regulated in gliomas tissues compared with normal brain tissues, which was negatively correlated with the stage of gliomas. In addition, miR-15b was significantly down-regulated in GSCs compared with non-GSCs. Up-regulation of miR-15b significantly reduced the migration and invasion ability of GSCs (P < 0.01), and down-regulation of miR-15b significantly enhanced the cell migration and invasion of GSCs (P < 0.01). By target prediction analysis, we obtained that ABCG2 was a potential target gene of miR-15b. Luciferase assay confirmed that miR-15b targeted ABCG2 directly, and migration and invasion of GSCs were dramatically reduced by ABCG2 siRNA (P < 0.01). ELISA results showed that up-regulation miR-15b significantly inhibited MMP-2/-9 expression. ELISA and gelatin zymography assay results showed that ABCG2 siRNA did not affect MMP-2/-9 expression, but significantly inhibited the activity of MMP-2/-9. In the in vivo tumor model, GSCs were more tumorigenic as compared with non-GSCs from the same tumor in vivo. To conclude, miR-15b regulates the migration and invasion of GSCs by targeting the ABCG2 signaling pathway, and up-regulation of miR-15b can suppress the migration and invasion of GSCs.
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Objective To analyze the risk factors of repeat revascularization after percutaneous coronary intervention (PCI) in elderly patients.Methods A total of 315 patients,aged 60-99 (72.89±7.80) years hospitalized during 2004 and 2015 due to coronary heart disease and received PCI therapy in the geriatric department of Peking University First Hospital were enrolled.Patients who received repeat revascularization were divided into the target lesion repeat revascularization (TLR)group and the non target lesion revascularization (non-TLR)group.Multivariate regression analysis was performed to make comparisons with non repeat revascularization group respectively and evaluate related risk factors.The median follow-up was 62months.Results The rate of TLR was less than non-TLR in elderly patients.Multivariate regression analysis showed that diabetes mellitus (HR 4.56,95% CI 1.94-10.75,P<0.05) and number of stents (HR 1.55,95% CI 1.05-2.29,P<0.05) were related risk factor of TLR,while age (HR 0.94,95% CI 0.90-0.98,P<0.05),the value of LDL-C reduction (HR 2.51,95% CI 1.56-3.99,P<0.05),the proportion of bifurcation lesions (HR 2.24,95% CI 1.20-4.17,P<0.05) and residual SYNTAX score (rSS) (HR 1.07,CI95% 1.02-1.11,P<0.05) wererelated risk factors of non-TLI.Conclusions The incidence of non-TLR were higher than TLR in elderly patients.Increased total number of stent implantation and diabetes mellitus were related risk factors of TLR,while the lower average age,less reduction of LDL-C,increased proportion of bifurcation lesions and higher rSS were related risk factors of non-TLR.
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Objective To investigate ADP-induced platelet aggregation rate(ADP-Ag),safety and efficacy of different ticagrelor dosage in elderly patients undergoing PCI. Methods 48 elderly patients aged 60 or older were enrolled. After PCI treatment, the patients received antiplatelet therapy with ticagrelor 90 mg(n=26)or 45 mg (n=22)twice daily. ADP-Ag was measured on day 3 to 7 after initial ticagrelor therapy and compared between the two different-dose ticagrelor groups. ADP-Ag ticagrelor treatment was also compared to measurement if patients were previously taking clopidogrel 75 mg daily. Major adverse cardiovascular events (MACE)and bleeding events were recorded in following 12 months. Results Inhibition of ADP-induced platelet aggregation was greater for ticagrelor 90 mg BID versus 45 mg BID(ADP-Ag(27.88±7.77)% vs.(37.87±2.90)%,P<0.05). During the follow-up period,no gastrointestinal bleedings,cerebral hemorrhage or thrombus events occurred in all patients. In ticagrelor 90 mg BID group,stent-restenosis occurred in 3 patients and they needed to take revascularization therapy. Minimal bleeding events occurred in 4 patients(15.4%)and 1 patient(4.5%) with ticagrelor 90 mg and 45 mg BID treatment,respectively. The ADP-Ag with clopidogrel 75 mg QD treatment previously was(51.18±5.55)%,and declined to(26.87±7.33)% and(38.29±2.65)% after conversion to ticagrelor 90 mg BID and 45 mg BID treatment,respectively. Conclusions Ticagrelor of 90 mg BID or 45 mg BID both inhibited ADP-induced platelet aggregation better than clopidogrel. Ticagrelor 45 mg BID was not inferior to 90 mg BID in preventing MACE and with less minimal bleeding events in elderly patients undergoing PCI.
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This study was designed to compare the curative effect, prognosis and safety of different preconditioning regimens for patients who received autologous hematopoietic stem cell transplantation (AHSCT) for malignant lymphoma (ML). The clinical data of 100 ML patients (Sep 1992 to Aug 2010 in 307 Hospital) were retrospectively analyzed, and were divided into two groups by different preconditioning regimens: the high-dose chemotherapy preconditioning group and high-dose chemotherapy/radiotherapy preconditioning group. The overall survival (OS) rate, progress free survival (PFS) rate and adverse effect were analyzed. The results showed that until Feb 2011, the median follow-up was 33.5 months. All patients were engrafted and their hematopoiesis was reconstituted. The median time of WBC recovery up to > 1.0×1.0(9)/L in high-dose chemotherapy preconditioning group and high-dose chemotherapy/radiotherapy preconditioning group were (6.0 ± 0.4) d and (8.2 ± 0.4) d, platelet up to > 20.0×1.0(9)/L in two groups were (7.1 ± 0.8) d and (11.4 ± 2.5) d (P < 0.05). The 3-year OS rate of the two groups were 67.3% and 68.9%. 5-year OS rates of two groups were 62.8% and 60.6%, 10-year OS rates of two groups were 57.6% and 56.2% respectively; 3-year PFS of two group were 63.6% and 63.2%, 5-year of two group were 59.4% and 58.3%, 10-year of two group were 50.8% and 55.3% respectively (P > 0.05). Meanwhile, the incidence of fever, infection, bleeding, secondary cancer between two groups was not significant different (P > 0.05). It is concluded that the hematopoietic reconstitution of high-dose chemotherapy/radiotherapy preconditioning group is later than that of high-dose chemotherapy preconditioning group. However, there is no significant difference in curative effect and prognosis between the two groups.
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Adolescente , Adulto , Idoso , Criança , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Adulto Jovem , Terapia Combinada , Transplante de Células-Tronco Hematopoéticas , Linfoma , Cirurgia Geral , Terapêutica , Prognóstico , Estudos Retrospectivos , Condicionamento Pré-Transplante , Métodos , Transplante AutólogoRESUMO
This paper explored the curative effect of combined modality therapy and extended field radiotherapy for early-stage Hodgkin's Lymphoma. 104 cases of early-stage Hodgkin's Lymphoma from Jan 1987 to Dec 2010 in PLA Hospital 307 were retrospectively analyzed, including 76 cases in combined modality therapy group and 28 cases in extended field radiotherapy group, and the long-term efficacy and toxicity of two therapy modalities were evaluated. The results showed that the median survival time of 104 cases was 85.42 months, the complete remission rates of combined modality therapy and extended field radiotherapy groups were 72.4 and 71.4 respectively (P = 0.924); the overall response rates of combined modality therapy and extended field radiotherapy groups were 97.4 and 96.4 respectively (P = 0.779); the 5-year overall survival (OS) rates in the 2 groups were 89.5 and 89.1 respectively, and the 8-year OS rates of the 2 groups were 81.3 and 70.6. No statistical difference was found in above-mentioned 2 groups. Moreover, the 5-year progression free survival (PFS) rates of these 2 groups were 84.2 and 69.0 (P = 0.04), and 8-year PFS rates of these 2 groups were 80.0 and 55.5 (P = 0.04) respectively, the 5-year relapse rates of these 2 groups were 28.1 and 45.6 (P = 0.023) respectively. It is concluded that the combined modality therapy can raise the PFS rate and reduce the relapse rate as compared with extended field radiotherapy for early-stage Hodgkin's Lymphoma, but there is no difference in the overall survival rate between the 2 groups.
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Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Criança , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Adulto Jovem , Protocolos de Quimioterapia Combinada Antineoplásica , Terapia Combinada , Doença de Hodgkin , Tratamento Farmacológico , Radioterapia , Estudos Retrospectivos , Taxa de Sobrevida , Resultado do TratamentoRESUMO
This study was aimed to investigate the differences of therapeutic efficiencies, side effects and recovery rates of immune function in refractory lymphoma patients treated with autologous peripheral blood hematopoietic stem cell transplantation (APBHSCT), autologous bone marrow transplantation (ABMT) and combination of APBHSCT with ABMT (APBHSCT + ABMT) by retrospective analysis, and to evaluate the merits and demerits of 3 kinds of transplantation for treatment of refractory lymphoma. 68 patients with malignant lymphoma were treated with autologous hematopoietic stem cells transplantation. Out of 68 patients 10 cases were treated with autologous bone marrow transplantation (ABMT), 46 cases were treated with autologous peripheral blood hematopoietic stem cell transplantation (APBHSCT), and 12 cases were treated with autologous peripheral blood hematopoietic stem cells transplantation combined with autologous bone marrow transplantation (APBHSCT + ABMT). The results indicated that the therapeutic response rates and survival rates at 1, 3, 5 years for each transplant regimen were 90% and 75%, 57.1%, 33.3%; 86.4% and 74.4%, 54.2%, 38.1%; 83.3% and 72.7%, 55.6%, 40%. The times of ANC > or = 0.5 x 10(9)/L were 13, 11 and 9 days, times of platelet >/= 20 x 10(9)/L were 17, 14 and 10 days. The recovery rates of T cell subtypes in patients received ABMT, APBHSCT and APBHSCT + ABMT on 3 months, 6 months, 1 year were (0%, 33.3%, 60%), (10.8%, 32%, 73.9%), (27.3%, 55.6%, 85.7%) respectively. In conclusion, the efficacy and side effects of APBHSCT + ABMT as compared with ABMT and APBHSCT are roughly the same, but ABMT + APBHSCT can result in more rapid hematopoietic reconstitution and less restrictions with contributes to widen choice of transplant regimen for patients with alder age and impaired hematopoietic functions.
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Adolescente , Adulto , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Adulto Jovem , Transplante de Medula Óssea , Linfoma , Cirurgia Geral , Transplante de Células-Tronco de Sangue Periférico , Estudos Retrospectivos , Transplante AutólogoRESUMO
Sphingolipids are structural components of cellular membranes.Their metabolites,such as ceramide,sphingosine,and sphingosine 1-phosphate are important bioactive molecules,which act as the first or second messengers regulating various cellular activities,including proliferation,survival,migration and neovascularization in vitro and in vivo tumor models.Sphingosine kinases 1(SPK1) is a critical regulator of the balance between ceramide and S1P.Recent studies show that ceramide,SPK1 and S1P can regulate many of the hallmarks of cancer.These studies suggest that these metabolites can serve as novel targets for cancer therapy.
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Natural phytoestrogens such as the isoflavones genistein and daidzein, and the flavones quercetin exhibit anti-cancer properties. This study was purpose to investigate the anti-proliferative effect of phytoestrogens on acute myeloid leukemia (AML) and chronic myeloid leukemia (CML) cells, and their synergistic antileukemic effect in combination with chemotherapeutic drugs. Optimal dosage of genistein, quercetin and in combination with chemicals for leukemia cells were determined by experiments. Cell viability, apoptosis induction and cell cycle arrest were detected by trypan blue staining, MTT assay, optical microscopy, flow cytometry (FCM). The schedule treatment of combination of genistein and chemicals was determined. The results showed that genistein exhibited a dose- and time-dependent inhibitory effect on cell proliferation in NB4 and HL-60 cells, induced apoptosis and cell cycle arrest in G2/M phase. Quercetin had evident inhibitory effect on the proliferation of K562 and K562/A cells. The combination of genistein and chemicals exerted a synergistic effect on cell growth inhibition. In conclusion, this study demonstrated the synergistic antileukemic effect of genistein with chemotherapeutic drugs on leukemic cells. This combination appears to be a new idea for the clinical novel treatment of leukemia.
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Humanos , Antineoplásicos , Farmacologia , Apoptose , Proliferação de Células , Sinergismo Farmacológico , Genisteína , Farmacologia , Células HL-60 , Isoflavonas , Farmacologia , Leucemia Monocítica Aguda , Patologia , Leucemia Mieloide Aguda , Patologia , Fitoestrógenos , Farmacologia , Quercetina , FarmacologiaRESUMO
Tumor angiogenesis is a complicated process in which VEGF and the members of Ang family have been proposed to play an important role. Ang family is the only vascular growth factor family including activators and inhibitors. They can all bind with specific tyrosine kinase receptor Tie-2 and participate in angiogenesis. Vascular endothelial growth factor (VEGF) is secreted by tumor cells and coordinates with Ang to promote angiogenesis. Further research on the structure and functional mechanism of VEGF and Ang will contribute to develop a new type of antiangiogenesis medicine, which might have good perspect in clinical practice. In this paper, the structure and action mechanism of Ang family and its receptor Tie-2, the application of Ang family and Tie in tumor therapy, and the synergic mechanism between Ang and VEGF were summarized.
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Animais , Humanos , Inibidores da Angiogênese , Farmacologia , Angiopoietina-2 , Farmacologia , Sinergismo Farmacológico , Neoplasias , Neovascularização Patológica , Fatores de Crescimento do Endotélio Vascular , FarmacologiaRESUMO
<p><b>OBJECTIVE</b>To determine the clinical results of selected CD34(+) cell autologous transplantation in advanced malignant tumors.</p><p><b>METHODS</b>After pretreatment, fifteen patients aged 12 - 70 (49.5) years with various Stage III or IV malignant tumors were given the sorted CD34(+) cells collected by magnetic-activated cell sorting (Clini MACS, Milteny Biotech, Germany).</p><p><b>RESULTS</b>Peripheral blood progenitor cells (PBPC) from the patients were mobilized by chemotherapy and G-CSF 5 micro g/kg per day. CD34(+) cells gave 2.0 - 5 log depletion after cell sorting, with a median yield of CD34(+) selected cells of 2.4 (0.15 - 12.03) x 10(6)/kg. It gave a median recovery of 64 (52 - 81.4)% and median purity of 98.2 (83.2 - 99.7)%. The median time of neutrophil recovery > 1.0 x 10(9)/L and platelet recovery > 20 x 10(9)/L post-transplantation were 14 (8 - 26) days and 13 (11 - 35) days, respectively. On follow-up of 2 - 33 (11) months, the event-free survival rate was 53.3% (8/15) and the overall survival rate was 66.7% (10/15).</p><p><b>CONCLUSION</b>Transplantation of autologous selected PBPC CD34(+) cells gives prompt and stable engraftment. Selected CD34(+) cell transplantation, being a safe approach, may improve the clinical outcome even in patients with advanced malignant tumors.</p>
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Adolescente , Adulto , Idoso , Criança , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Antígenos CD34 , Transplante de Células-Tronco Hematopoéticas , Neoplasias , Mortalidade , Terapêutica , Taxa de Sobrevida , Transplante AutólogoRESUMO
The purpose of this study was to evaluate whether the DNA vaccine containing idiotypic gene fragment of human B-cell lymphoma cell line Namalwa could elicit the specific anti-idiotypic immune response in vivo. The candidate gene fragment of the lymphoma cell, variable region of heavy chain (VH) of the membranous immunoglobulin, was amplified using Ig superfamily primers by means of RT-PCR. Also, the intact cDNA of murine monocyte chemoattractant protein (MCP-3) was cloned and used as the adjuvant molecular. The two gene fragments of VH and MCP-3 were fused together by 8aa linker peptide with recombinant PCR. Subsequently, the fusion gene fragment was cloned into eukaryonic expression vector pcDNA3.1 to construct DNA vaccine plasmid. Prior to the immunization, the transient transfection coupled with RT-PCR was performed to prove that the recombinant plasmid could express in eukaryonic cells in right way. Then two groups of mice were immunized by intramuscular injection with DNA vaccine and mock plasmid pcDNA3.1 respectively. Three times of injection were performed with 100 micro g plasmid respectively at the beginning of the experiment and 2, 4 weeks after the first injection for all the groups. FACS analysis was chosen to detect the antibodies recognizing lymphoma cells at different time following vaccination. The results demonstrated that specific anti-idiotypic antibody could be detected in the group of DNA vaccine immunized mice as early as eight weeks after the first immunization. Further test demonstrated that the anti-idiotypic antibody could maintain for at least twenty weeks with high titer. Anti-idiotypic antibodies were elicited in three of five mice of the DNA vaccine immunized group. The Abs of DNA vaccine immunized mice could only recognize Namalwa cell line instead of another unrelated human cell line A549. There is no cellular response detected in the DNA vaccine immunized mice. It is concluded that the DNA vaccine containing fused MCP3-VH sequence could elicit specific anti-idiotypic antibody against B-cell lymphoma in vivo and could be used in further study of DNA vaccine against B-cell lymphoma. The results would provide the basis for further studies and optimization of this therapeutic strategy on patients with B-lymphoproliferative disease.
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Animais , Humanos , Camundongos , Anticorpos Anti-Idiotípicos , Sangue , Células COS , Vacinas Anticâncer , Alergia e Imunologia , Quimiocina CCL7 , Citocinas , Imunização , Cadeias Pesadas de Imunoglobulinas , Genética , Região Variável de Imunoglobulina , Genética , Linfoma de Células B , Alergia e Imunologia , Camundongos Endogâmicos BALB C , Proteínas Quimioatraentes de Monócitos , Genética , Alergia e Imunologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Vacinas de DNA , Alergia e ImunologiaRESUMO
The lymphochip is a special DNA chip which is used to monitor the gene expression profiling of malignant lymphoma. The basic principle, technological procedure and types of lymphochip were briefly reviewed in the article. The article emphatically introduced the advance of lymphochip implication which was used to identify the molecular diagnosis, distinguish the subtypes and research the morbific mechanism of malignant lymphoma. The aspects of development and problems in lymphochip study were discussed.